| 1. |
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| 2. |
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| 3. |
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| 4. |
- Muller, D, et al.
(författare)
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Competitive adsorption of gelatin and sodium dodecylbenzenesulfonate at hydrophobic surfaces
- 1998
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Ingår i: Langmuir. - 0743-7463 .- 1520-5827. ; 14, s. 3107-3114
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Tidskriftsartikel (refereegranskat)abstract
- The competitive adsorption of gelatin and sodium dodecylbenzenesulfonate (SDBS) at hydrophobic surfaces was investigated with surface and interfacial tension measurements, ellipsometry, surface plasmon resonance spectroscopy (SPR), and total internal reflectance fluorescence spectroscopy (TIRF). From both ellipsometry and SPR, initial additions of SDBS after gelatin preadsorption were found to result in a total adsorbed amount increase, as well as in a swelling of the adsorbed layer. At higher SDBS concentrations, both the total adsorbed amount and the amount of gelatin adsorbed decrease, which was observed from ellipsometry, SPR, and TIRF. From surface and interfacial tension measurements, it was found that the critical aggregation concentration (cac) for the SDBS-gelatin system decreases with decreasing pH. Analogous to this, ellipsometry, SPR, and TIRF indicate that the SDBS concentration required to cause a significant decrease in the gelatin adsorbed amount decreases with decreasing pH. The desorption therefore seems to be correlated to the SDBS binding to the adsorbed gelatin molecules rather than to purely competitive adsorption.
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| 5. |
- Muller, D, et al.
(författare)
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Interaction of gelatin and sodium dodecyl benzene sulphonate at oil and water interfaces
- 1997
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Ingår i: Imaging Science Journal. - 1368-2199 .- 1743-131X. ; 45, s. 229-235
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Tidskriftsartikel (refereegranskat)abstract
- Interaction between sodium dodecyl benzene sulphonate (SDBS) and gelatin was studied in relation to emulsification behaviour and emulsion stability. We chose two different oils to study influences of oil phase characteristics, namely, tricresyl phosphate (TCP) as an oil with polar and slightly hydrophilic nature, and n-docedane (nC12) as its apolar contrast. Our interfacial tension measurements showed that both TCP and n-C12 give critical values (i.e., cac and cmc) very close to those of surface tension measurement. This result indicates that the complexation behaviour in bulk solution is independent of the presence or the nature of the oil phase. Absolute tension values above the cmc and slope values at the cmc in gelatin free systems, however, imply SDBC's weaker adsorption to TCP than to n-C12. Our emulsification results for the TCP system revealed the existence of an optiomal point for emulsion stability in the SDBS concentration region between the cac and the cmc. Above this point, emulsion stability deteriorates remarkably. The behaviour is in line with our findings from the dynamic sorption expriments (ellipsometry, TIRF, and SPR) reported elsewhere, which showed a rapid desorption of gelatin from the hydrophobic surfaces above the cac. The results suggest that the adsorption of gelatin/ surfactant complexes at the interface is a key factor for stability of the polar oil emulsion system. Contrary to that , the n-c12 system did not show any deterioration, even above the cmc, which is presumably due to a strong double layer effect from the firmly adsorbed layer of SDBS at the interface.
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| 6. |
- Petrlova, Jitka, et al.
(författare)
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Aggregation of thrombin-derived C-terminal fragments as a previously undisclosed host defense mechanism
- 2017
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 114:21, s. E4213-E4222
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Tidskriftsartikel (refereegranskat)abstract
- Effective control of endotoxins and bacteria is crucial for normal wound healing. During injury, the key enzyme thrombin is formed, leading to generation of fibrin. Here, we show that human neutrophil elastase cleaves thrombin, generating 11-kDa thrombin-derived C-terminal peptides (TCPs), which bind to and form amorphous amyloid-like aggregates with both bacterial lipopolysaccharide (LPS) and gram-negative bacteria. In silico molecular modeling using atomic resolution and coarse-grained simulations corroborates our experimental observations, altogether indicating increased aggregation through LPS-mediated intermolecular contacts between clusters of TCP molecules. Upon bacterial aggregation, recombinantly produced TCPs induce permeabilization of Escherichia coli and phagocytic uptake. TCPs of about 11 kDa are present in acute wound fluids as well as in fibrin sloughs from patients with infected wounds. We noted aggregation and colocalization of LPS with TCPs in such fibrin material, which indicates the presence of TCP-LPS aggregates under physiological conditions. Apart from identifying a function of proteolyzed thrombin and its fragments, our findings provide an interesting link between the coagulation system, innate immunity, LPS scavenging, and protein aggregation/amyloid formation.
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| 7. |
- Bartholeyns, J, et al.
(författare)
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Cellular vaccines
- 1998
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Ingår i: Research in Immunology. - 0923-2494 .- 1879-1425. ; 149, s. 647-649
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Tidskriftsartikel (refereegranskat)abstract
- This project is devoted to the development of novel cellular vaccines designed to treat cancer patients. These cellular vaccines present and enhance immunogens, which will elicit a potent immune response. The goal is to achieve safe and effective immune reaction against the patient's own tumour. (1) Autologous cellular vaccines are prepared by processing circulating brood mononuclear cells outside of the patient's body (ex vivo) to differentiate them into antigen-presenting cells (APCs). Monocyte-derived APCs (MD-APCs) are then grown in the presence of exogenous target antigens (tumour cell debris, or apoptotic bodies) to become fully mature APCs. (2) Functionality for antigen presentation to T cells of ex vivo MD-APCs is evaluated in vivo. (3) Cellular vaccines are tested in selected rodent animal models. Efficiency and immune response are monitored in pertinent experimental systems for cancer. Pharmacological data are generated for clinical investigation. Tolerance and biologic effects are documented in primates. (4) The first clinical trials on cancer patients are taking place in 1998 on melanoma and prostate cancer to validate the concept. Specialized eel processors with dedicated software and standardized controls are being developed and used for the preparation of cellular vaccines. (5) The evaluation of new non-viral vectors and the validation of new non-viral transfection methods of mononuclear cells with marker genes is in progress and will lead to the ex vivo transfection of genes coding for immunostimulating cytokines or for tumour antigens in MD-APCs. Efficiency will be validated in vitro and in animal models. The ex vivo and animal model studies validate the clinical relevance of this new cellular immunotechnology. Clinical validation of individual autologous cellular vaccines in specific indications for which no treatment is presently available will allow the development of cellular and gene immunotherapy for other types of cancers.
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| 8. |
- Caselli, Lucrezia, et al.
(författare)
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Boosting Membrane Interactions and Antimicrobial Effects of Photocatalytic Titanium Dioxide Nanoparticles by Peptide Coating
- 2024
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Ingår i: Small. - : John Wiley and Sons Inc. - 1613-6810 .- 1613-6829.
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Tidskriftsartikel (refereegranskat)abstract
- Photocatalytic nanoparticles offer antimicrobial effects under illumination due to the formation of reactive oxygen species (ROS), capable of degrading bacterial membranes. ROS may, however, also degrade human cell membranes and trigger toxicity. Since antimicrobial peptides (AMPs) may display excellent selectivity between human cells and bacteria, these may offer opportunities to effectively “target” nanoparticles to bacterial membranes for increased selectivity. Investigating this, photocatalytic TiO2 nanoparticles (NPs) are coated with the AMP LL-37, and ROS generation is found by C11-BODIPY to be essentially unaffected after AMP coating. Furthermore, peptide-coated TiO2 NPs retain their positive ζ-potential also after 1–2 h of UV illumination, showing peptide degradation to be sufficiently limited to allow peptide-mediated targeting. In line with this, quartz crystal microbalance measurements show peptide coating to promote membrane binding of TiO2 NPs, particularly so for bacteria-like anionic and cholesterol-void membranes. As a result, membrane degradation during illumination is strongly promoted for such membranes, but not so for mammalian-like membranes. The mechanisms of these effects are elucidated by neutron reflectometry. Analogously, LL-37 coating promoted membrane rupture by TiO2 NPs for Gram-negative and Gram-positive bacteria, but not for human monocytes. These findings demonstrate that AMP coating may selectively boost the antimicrobial effects of photocatalytic NPs. © 2024 The Authors.
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| 9. |
- Eriksson, J, et al.
(författare)
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Model cellulose films exposed to H. insolens glucoside hydrolase family 45 endo-cellulase—the effect of the carbohydrate-binding module
- 2005
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 285, s. 94-99
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Tidskriftsartikel (refereegranskat)abstract
- The effects of enzyme structure and activity on the degradation of model cellulose substrates were investigated by ellipsometry for the cellulase Humicola insolens GH45. The inactive variant D10N was found to adsorb at the cellulose surface but also to be incorporated into the cellulose films to an extent that depended on pH. For the native protein, the initial adsorption monitored for the inactive variant D10N was followed by enzyme-mediated degradation of the cellulose films. Again, a dependence on pH was found, such that higher pH resulted in slower enzymatic degradation. Removing the carbohydrate-binding module eliminated this pH dependence but also resulted in a decreased adsorption to the cellulose surface, and in a decreased net catalytic effect
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| 10. |
- Griffiths, PC, et al.
(författare)
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Role of copolymer architecture on adsorption at the solid/liquid interface
- 1998
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Ingår i: Langmuir. - 0743-7463 .- 1520-5827. ; 14, s. 1779-1785
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Tidskriftsartikel (refereegranskat)abstract
- The adsorption of monodisperse block copolymers comprising poly(ethylene oxide)-poly(butylene oxide) onto polystyrene latex from aqueous solution has been investigated by small-angle neutron scattering and photon correlation spectroscopy with particular reference to the role of molecular architecture. It appears that chain architecture is (i) a weak factor in the adsorption behavior when the hydrophobic block is located in the center of the polymer, since the triblock E100B15E100 behaved very similarly to the cyclic c-E200B15, but (ii) a significant factor when the hydrophobic block is located at the end of the copolymer chain, as shown by the more dense and thicker layer formed by E200B15 compared to the triblock E100B15E100. The hydrodynamic thickness of the layer formed by the small diblock E100B15 was approximately half that exhibited by the larger diblock E200B15. Good agreement was observed between depletion and SANS-derived adsorbed amounts. Theoretical predictions and self-consistent mean-field calculations of the adsorption also show excellent qualitative agreement with experiment.
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| 11. |
- Habibovic, Azra, et al.
(författare)
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Evaluating interactions with non-existing automated vehicles: three Wizard of Oz approaches
- 2016
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Ingår i: 2016 IEEE Intelligent Vehicles Symposium (IV). - Piscataway, NJ : IEEE. - 9781509018215 - 9781509018222 ; , s. 32-37
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Konferensbidrag (refereegranskat)abstract
- Highly automated test vehicles are rare today, and (independent) researchers have often limited access to them. Also, developing fully functioning system prototypes is time and effort consuming. In this paper, we present three adaptions of the Wizard of Oz technique as a means of gathering data about interactions with highly automated vehicles in early development phases. Two of them address interactions between drivers and highly automated vehicles, while the third one is adapted to address interactions between pedestrians and highly automated vehicles. The focus is on the experimental methodology adaptations and our lessons learned.
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| 12. |
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| 13. |
- Holdbrook, Daniel A., et al.
(författare)
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Influence of pH on the activity of thrombin-derived antimicrobial peptides
- 2018
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Ingår i: Biochimica et Biophysica Acta - Biomembranes. - : Elsevier BV. - 0005-2736 .- 1879-2642. ; 1860:11, s. 2374-2384
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Tidskriftsartikel (refereegranskat)abstract
- The wound environment is characterized by physiological pH changes. Proteolysis of thrombin by wound-derived proteases, such as neutrophil elastase, generates antimicrobial thrombin-derived C-terminal peptides (TCPs), such as HVF18 (HVFRLKKWIQKVIDQFGE). Presence of such TCPs in human wound fluids in vivo, as well as the occurrence of an evolutionarily conserved His residue in the primary amino acid sequence of TCPs, prompted us to investigate the pH-dependent antibacterial action of HVF18, as well as of the prototypic GKY25 (GKYGFYTHVFRLKKWIQKVIDQFGE). We show that protonation of this His residue at pH 5.5 increases the antibacterial activity of both TCPs against Gram-negative Escherichia coli by membrane disruption. Physiological salt level (150 mM NaCl) augments antibacterial activity of GKY25 but diminishes for the shorter HVF18. Replacing His with Leu or Ser in GKY25 abolishes the His protonation-dependent increase in antibacterial activity at pH 5.5, whereas substitution with Lys maintains activity at neutral (pH 7.4) and acidic pH. Interestingly, both TCPs display decreased binding affinities to human CD14 with decreasing pH, suggesting a likely switch in mode-of-action, from anti-inflammatory at neutral pH to antibacterial at acidic pH. Together, the results demonstrate that apart from structural prerequisites such as peptide length, charge, and hydrophobicity, the evolutionarily conserved His residue of TCPs influences their antibacterial effects and reveals a previously unknown aspect of TCPs biological action.
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| 14. |
- Ilyas, Humaira, et al.
(författare)
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Effect of PEGylation on Host Defense Peptide Complexation with Bacterial Lipopolysaccharide
- 2021
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Ingår i: Bioconjugate Chemistry. - : American Chemical Society (ACS). - 1043-1802 .- 1520-4812. ; 32:8, s. 1729-1741
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Tidskriftsartikel (refereegranskat)abstract
- Conjugation with poly(ethylene glycol) ("PEGylation") is a widely used approach for improving the therapeutic propensities of peptide and protein drugs through prolonging bloodstream circulation, reducing toxicity and immunogenicity, and improving proteolytic stability. In the present study, we investigate how PEGylation affects the interaction of host defense peptides (HDPs) with bacterial lipopolysaccharide (LPS) as well as HDP suppression of LPS-induced cell activation. In particular, we investigate the effects of PEGylation site for KYE28 (KYEITTIHNLFRKLTHRLFRRNFGYTLR), a peptide displaying potent anti-inflammatory effects, primarily provided by its N-terminal part. PEGylation was performed either in the N-terminus, the C-terminus, or in both termini, keeping the total number of ethylene groups (n = 48) constant. Ellipsometry showed KYE28 to exhibit pronounced affinity to both LPS and its hydrophobic lipid A moiety. The PEGylated peptide variants displayed lower, but comparable, affinity for both LPS and lipid A, irrespective of the PEGylation site. Furthermore, both KYE28 and its PEGylated variants triggered LPS aggregate disruption. To investigate the peptide structure in such LPS complexes, a battery of nuclear magnetic resonance (NMR) methods was employed. From this, it was found that KYE28 formed a well-folded structure after LPS binding, stabilized by hydrophobic domains involving aromatic amino acids as well as by electrostatic interactions. In contrast, the PEGylated peptide variants displayed a less well-defined secondary structure, suggesting weaker LPS interactions in line with the ellipsometry findings. Nevertheless, the N-terminal part of KYE28 retained helix formation after PEGylation, irrespective of the conjugation site. For THP1-Xblue-CD14 reporter cells, KYE28 displayed potent suppression of LPS activation at simultaneously low cell toxicity. Interestingly, the PEGylated KYE28 variants displayed similar or improved suppression of LPS-induced cell activation, implying the underlying key role of the largely retained helical structure close to the N-terminus, irrespective of PEGylation site. Taken together, the results show that PEGylation of HDPs can be done insensitively to the conjugation site without losing anti-inflammatory effects, even for peptides inducing such effects through one of its termini.
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| 15. |
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| 16. |
- Lilja, AM, et al.
(författare)
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Neural Stem Cell Transplant-Induced Effect on Neurogenesis and Cognition in Alzheimer Tg2576 Mice Is Inhibited by Concomitant Treatment with Amyloid-Lowering or Cholinergic α7 Nicotinic Receptor Drugs
- 2015
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Ingår i: Neural plasticity. - : Hindawi Limited. - 1687-5443 .- 2090-5904. ; 2015, s. 370432-
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Tidskriftsartikel (refereegranskat)abstract
- Stimulating regeneration in the brain has the potential to rescue neuronal networks and counteract progressive pathological changes in Alzheimer’s disease (AD). This study investigated whether drugs with different mechanisms of action could enhance neurogenesis and improve cognition in mice receiving human neural stem cell (hNSC) transplants. Six- to nine-month-old AD Tg2576 mice were treated for five weeks with the amyloid-modulatory and neurotrophic drug (+)-phenserine or with the partialα7 nicotinic receptor (nAChR) agonist JN403, combined with bilateral intrahippocampal hNSC transplantation. We observed improved spatial memory in hNSC-transplanted non-drug-treated Tg2576 mice but not in those receiving drugs, and this was accompanied by an increased number of Doublecortin- (DCX-) positive cells in the dentate gyrus, a surrogate marker for newly generated neurons. Treatment with (+)-phenserine did however improve graft survival in the hippocampus. An accumulation ofα7 nAChR-expressing astrocytes was observed around the injection site, suggesting their involvement in repair and scarring processes. Interestingly, JN403 treatment decreased the number ofα7 nAChR-expressing astrocytes, correlating with a reduction in the number of DCX-positive cells in the dentate gyrus. We conclude that transplanting hNSCs enhances endogenous neurogenesis and prevents further cognitive deterioration in Tg2576 mice, while simultaneous treatments with (+)-phenserine or JN403 result in countertherapeutic effects.
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| 17. |
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| 18. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Interaction of Laponite with Membrane Components - Consequences for Bacterial Aggregation and Infection Confinement
- 2019
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Ingår i: ACS Applied Materials and Interfaces. - : American Chemical Society (ACS). - 1944-8244 .- 1944-8252. ; 11:17, s. 15389-15400
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Tidskriftsartikel (refereegranskat)abstract
- The antimicrobial effects of Laponite nanoparticles with or without loading of the antimicrobial peptide LL-37 was investigated along with their membrane interactions. The study combines data from ellipsometry, circular dichroism, fluorescence spectroscopy, particle size/ζ potential measurements, and confocal microscopy. As a result of the net negative charge of Laponite, loading of net positively charged LL-37 increases with increasing pH. The peptide was found to bind primarily to the outer surface of the Laponite nanoparticles in a predominantly helical conformation, leading to charge reversal. Despite their net positive charge, peptide-loaded Laponite nanoparticles did not kill Gram-negative Escherichia coli bacteria or disrupt anionic model liposomes. They did however cause bacteria flocculation, originating from the interaction of Laponite and bacterial lipopolysaccharide (LPS). Free LL-37, in contrast, is potently antimicrobial through membrane disruption but does not induce bacterial aggregation in the concentration range investigated. Through LL-37 loading of Laponite nanoparticles, the combined effects of bacterial flocculation and membrane lysis are observed. However, bacteria aggregation seems to be limited to Gram-negative bacteria as Laponite did not cause flocculation of Gram-positive Bacillus subtilis bacteria nor did it bind to lipoteichoic acid from bacterial envelopes. Taken together, the present investigation reports several novel phenomena by demonstrating that nanoparticle charge does not invariably control membrane destabilization and by identifying the ability of anionic Laponite nanoparticles to effectively flocculate Gram-negative bacteria through LPS binding. As demonstrated in cell experiments, such aggregation results in diminished LPS-induced cell activation, thus outlining a promising approach for confinement of infection and inflammation caused by such pathogens.
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| 19. |
- Malekkhaiat Häffner, Sara, et al.
(författare)
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Nanoclay-induced bacterial flocculation for infection confinement
- 2020
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier. - 0021-9797 .- 1095-7103. ; 562, s. 71-80
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Tidskriftsartikel (refereegranskat)abstract
- Effects of size and charge of anionic nanoclays on their interactions with bacteria-mimicking lipid membranes, bacterial lipopolysaccharide (LPS), and Gram-negative bacteria were investigated using ellipsometry, dynamic light scattering, ζ-potential measurements, and confocal microscopy combined with Live/Dead staining. Based on particle size and charge density, three different anionic hectorite nanoclays were employed, and investigated in the presence and absence of the net cationic human antimicrobial peptide LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES). In the absence of this peptide, the nanoclays were found not to bind to similarly anionic bacteria-mimicking model phospholipid membranes, nor to destabilize these. Similarly, while all nanoclays induced aggregation of Escherichia coli bacteria, the flocculated bacteria remained alive after aggregation. In contrast, LL-37 alone, i.e. in the absence of nanoclay particles, displays antimicrobial properties through membrane lysis, but does not cause bacterial aggregation in the concentration range investigated. After loading the nanoclays with LL-37, potent bacterial aggregation combined with bacterial membrane lysis was observed for all nanoclay sizes and charge densities. Demonstrating the potential of these combined systems for confinement of infection, LPS-induced NF-κB activation in human monocytes was found to be strongly suppressed after nanoclay-mediated aggregation, with a wide tolerance for nanoparticle size and charge density.
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| 20. |
- Malmsten, M, et al.
(författare)
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Adsorption of complement protein C3 at polymer surfaces
- 1996
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Ingår i: Journal of Colloid and Interface Science. - 0021-9797 .- 1095-7103. ; 179, s. 163-172
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Tidskriftsartikel (refereegranskat)abstract
- The adsorption of C3 at poly(methyl methacrylate) (PMMA) and poly(styrene) (PS) surfaces was investigated with in situ ellipsometry and compared to that at (hydrophilic and negatively charged) silica and (hydrophobic) methylated silica. The adsorption of C3 at PMMA was higher than that at PS, while the adsorbed layer thickness was the same for the two surfaces. For both PMMA and PS the adsorbed layer thickness (10±2 nm) corresponds rather closely to that of end-on oriented C3 molecules. The adsorption of C3 at PMMA and PS was found to be intermediate between that at silica and methylated silica, although the adsorbed layer thickness was similar for all surfaces. The competitive adsorption between C3, human serum albumin (HSA), and factor B was investigated with ellipsometry and total internal reflection fluorescence spectroscopy (TIRF). Addition of HSA after C3 preadsorption resulted in fractional C3 desorption for both PMMA and PS. Factor B deposition at PS after preadsorption of C3 and blocking with HSA was found to be largely due to specific binding to C3/C3b, while in the case of PMMA, factor B was largely accumulated through passive (displacement) adsorption.
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| 21. |
- Malmsten, M, et al.
(författare)
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Protein adsorption at n-butane plasma polymer surfaces
- 1996
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Ingår i: Colloids and Surfaces B. - 0927-7765 .- 1873-4367. ; 6, s. 191-199
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Tidskriftsartikel (refereegranskat)abstract
- The adsorption of human serum albumin (HSA) and fibrinogen at n-butane plasma polymer surfaces prepared by low temperature plasma polymerization at different energy inputs have been investigated with in situ ellipsometry. Within experimental uncertainty, the adsorption of both fibrinogen and HSA is constant over the power range used for the preparation of the n-butane surfaces and corresponds to that found for other hydrophobic surfaces at similar conditions. Furthermore, novel copolymer surfaces of n-butane and nitrogen at different ratios were prepared and investigated. Increasing the nitrogen content in the gas mixture during deposition resulted in an increased density of interfacial amine groups, as evidenced by an increased wettability, an increased interfacial nitrogen content, and an increased surface positive charge. This, in turn, was found to result in an increased fibrinogen adsorption, but in a weak decrease in the HSA adsorption.
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| 22. |
- Martin, Alma, et al.
(författare)
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In Situ Transformation of Electrospun Nanofibers into Nanofiber-Reinforced Hydrogels
- 2022
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Ingår i: Nanomaterials. - : MDPI AG. - 2079-4991. ; 12:14
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Tidskriftsartikel (refereegranskat)abstract
- Nanofiber-reinforced hydrogels have recently gained attention in biomedical engineering. Such three-dimensional scaffolds show the mechanical strength and toughness of fibers while benefiting from the cooling and absorbing properties of hydrogels as well as a large pore size, potentially aiding cell migration. While many of such systems are prepared by complicated processes where fibers are produced separately to later be embedded in a hydrogel, we here provide proof of concept for a one-step solution. In more detail, we produced core-shell nanofibers from the natural proteins zein and gelatin by coaxial electrospinning. Upon hydration, the nanofibers were capable of directly transforming into a nanofiber-reinforced hydrogel, where the nanofibrous structure was retained by the zein core, while the gelatin-based shell turned into a hydrogel matrix. Our nanofiber-hydrogel composite showed swelling to ~800% of its original volume and water uptake of up to ~2500% in weight. The physical integrity of the nanofiber-reinforced hydrogel was found to be significantly improved in comparison to a hydrogel system without nanofibers. Additionally, tetracycline hydrochloride was incorporated into the fibers as an antimicrobial agent, and antimicrobial activity against Staphylococcus aureus and Escherichia coli was confirmed.
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| 23. |
- Nordström, Randi, 1986-, et al.
(författare)
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Degradable dendritic nanogels as carriers for antimicrobial peptides
- 2019
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Ingår i: Journal of Colloid and Interface Science. - : Elsevier BV. - 0021-9797 .- 1095-7103. ; 554, s. 592-602
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, we investigate degradable anionic dendritic nanogels (DNG) as carriers for antimicrobial peptides (AMPs). In such systems, the dendritic part contains carboxylic acid-based anionic binding sites for cationic AMPs, whereas linear poly(ethylene glycol) (PEG) chains form a shell for promotion of biological stealth. In order to clarify factors influencing membrane interactions of such systems, we here address effects of nanogel charge, cross-linking, and degradation on peptide loading/release, as well as consequences of these factors for lipid membrane interactions and antimicrobial effects. The DNGs were found to bind the AMPs LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) and DPK-060 (GKHKNKGKKNGKHNGWKWWW). For the smaller DPK-060 peptide, loading was found to increase with increasing nanogel charge density. For the larger LL-37, on the other hand, peptide loading was largely insensitive to nanogel charge density. In line with this, results on the secondary structure, as well as on the absence of stabilization from proteolytic degradation by the nanogels, show that the larger LL-37 is unable to enter into the interior of the nanogels. While 40–60% nanogel degradation occurred over 10 days, promoted at high ionic strength and lower cross-linking density/higher anionic charge content, peptide release at physiological ionic strength was substantially faster, and membrane destabilization not relying on nanogel degradation. Ellipsometry and liposome leakage experiments showed both free peptide and peptide/DNG complexes to cause membrane destabilization, indicated also by antimicrobial activities being comparable for nanogel-bound and free peptide. Finally, the DNGs were demonstrated to display low toxicity towards erythrocytes even at peptide concentrations of 100 µM.
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| 24. |
- Nordström, Randi, et al.
(författare)
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Membrane interactions of microgels as carriers of antimicrobial peptides
- 2018
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Ingår i: Journal of Colloid and Interface Science. - : Academic Press Inc.. - 0021-9797 .- 1095-7103. ; 513, s. 141-150
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Tidskriftsartikel (refereegranskat)abstract
- Microgels are interesting as potential delivery systems for antimicrobial peptides. In order to elucidate membrane interactions of such systems, we here investigate effects of microgel charge density on antimicrobial peptide loading and release, as well as consequences of this for membrane interactions and antimicrobial effects, using ellipsometry, circular dichroism spectroscopy, nanoparticle tracking analysis, dynamic light scattering and z-potential measurements. Anionic poly(ethyl acrylate-co-methacrylic acid) microgels were found to incorporate considerable amounts of the cationic antimicrobial peptides LL-37 (LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) and DPK-060 (GKHKNKGKKNGKHNGWKWWW) and to protect incorporated peptides from degradation by infection-related proteases at high microgel charge density. As a result of their net negative z-potential also at high peptide loading, neither empty nor peptide-loaded microgels adsorb at supported bacteria-mimicking membranes. Instead, membrane disruption is mediated almost exclusively by peptide release. Mirroring this, antimicrobial effects against several clinically relevant bacteria (methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli, and Pseudomonas aeruginosa) were found to be promoted by factors facilitating peptide release, such as decreasing peptide length and decreasing microgel charge density. Microgels were further demonstrated to display low toxicity towards erythrocytes. Taken together, the results demonstrate some interesting opportunities for the use of microgels as delivery systems for antimicrobial peptides, but also highlight several key factors which need to be controlled for their successful use.
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| 25. |
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| 26. |
- Puthia, Manoj, et al.
(författare)
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Bioactive Suture with Added Innate Defense Functionality for the Reduction of Bacterial Infection and Inflammation
- 2023
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Ingår i: Advanced Healthcare Materials. - : Wiley. - 2192-2640 .- 2192-2659. ; 12:31, s. 1-20
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Tidskriftsartikel (refereegranskat)abstract
- Surgical site infections (SSI) are a clinical and economic burden. Suture-associated SSI may develop when bacteria colonize the suture surface and form biofilms that are resistant to antibiotics. Thrombin-derived C-terminal peptide (TCP)-25 is a host defense peptide with a unique dual mode of action that can target both bacteria and the excessive inflammation induced by bacterial products. The peptide demonstrates therapeutic potential in preclinical in vivo wound infection models. In this study, the authors set out to explore whether TCP-25 can provide a new bioactive innate immune feature to hydrophilic polyglactin sutures (Vicryl). Using a combination of biochemical, biophysical, antibacterial, biofilm, and anti-inflammatory assays in vitro, in silico molecular modeling studies, along with experimental infection and inflammation models in mice, a proof-of-concept that TCP-25 can provide Vicryl sutures with a previously undisclosed host defense capacity, that enables targeting of bacteria, biofilms, and the accompanying inflammatory response, is shown.
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| 27. |
- Saravanan, Rathi, et al.
(författare)
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Structural basis for endotoxin neutralisation and anti-inflammatory activity of thrombin-derived C-terminal peptides
- 2018
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Ingår i: Nature Communications. - : NATURE PUBLISHING GROUP. - 2041-1723. ; 9
-
Tidskriftsartikel (refereegranskat)abstract
- Thrombin-derived C-terminal peptides (TCPs) of about 2 kDa are present in wounds, where they exert anti-endotoxic functions. Employing a combination of nuclear magnetic resonance spectroscopy (NMR), biophysical, mass spectrometry and cellular studies combined with in silico multiscale modelling, we here determine the bound conformation of HVF18 (HVFRLKKWIQKVIDQFGE), a TCP generated by neutrophil elastase, in complex with bacterial lipopolysaccharide (LPS) and define a previously undisclosed interaction between TCPs and human CD14. Further, we show that TCPs bind to the LPS-binding hydrophobic pocket of CD14 and identify the peptide region crucial for TCP interaction with LPS and CD14. Taken together, our results demonstrate the role of structural transitions in LPS complex formation and CD14 interaction, providing a molecular explanation for the previously observed therapeutic effects of TCPs in experimental models of bacterial sepsis and endotoxin shock.
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| 28. |
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| 29. |
- Shao, Xing-Wu, et al.
(författare)
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Use of HIV-1 reverse transcriptase recovered from human plasma for phenotypic drug susceptibility testing
- 2003
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Ingår i: AIDS. ; 17, s. 1463-1471
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Tidskriftsartikel (refereegranskat)abstract
- Objective: To demonstrate the use of HIV-1 reverse transcriplase (RT) recovered directly fromplasma for phenotypic drug susceptibility testing. Methods: Plasma from HIV-1 infected individuals with and without drug resistance-associated mutations were selected for the study. The blind coded plasmas were treated to inactivate cellular enzymes. The virions were immobilized on a gel and washed to remove antiretroviral drugs and RT activity blocking antibodies. The immobilized virions were lysed; the viral RTeluted and quantified, all according to the ExaVir Load procedure. The drug sensitivity profiles of each RT were determined using serially diluted drugs and modified Cavidi HS Lenti RT kits. Results: The phenotypic drug sensitivity profiles of the RT and the patterns of drug resistance mutations were highly concordant. Plasma RT from virions devoid of mutations associated with drug resistance had average 50% inhibitory concentrations (IC50) of 1.5 +/- 0.93 muM for nevirapine, 0.21 +/- 0.099 muM forefavirenz, 7.1 +/- 3.2 muM for delavirdine, 0.42 +/- 0.15 muM for azidothymidine triphosphate and 0.059 +/- 0.018 muM for didehydrothymidine triphosphate. The increase in IC50 value for RT with drugresistance associated substitutions was from 3- to more than 65-fold for non-nucleoside inhibitors and between 2- and 30-fold for thymidine analogue drugs. Conclusion: RT derived from virions recovered from the plasma of HIV infected individuals can be used foranalysis of phenotypic drug susceptibility. The methods presented provide rapid alternatives for analysingphenotypic drug susceptibility especially when the therapy is based on non-nucleoside RT inhibitors and thymidine-analogue drugs.
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| 30. |
- Siegel, Günter, et al.
(författare)
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Atherogenesis and plaque rupture, surface/interface-related phenomena
- 2018
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Ingår i: Colloids and Surfaces A. - : ELSEVIER SCIENCE BV. - 0927-7757 .- 1873-4359. ; 557, s. 28-35
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Tidskriftsartikel (refereegranskat)abstract
- In atherogenesis, free oxygen radicals cause a lipid peroxidation of apoB100-containing lipoproteins in the blood, at the blood-endothelium-interface and in the subendothelial space. These lipoproteins easily aggregate, bind to their receptor heparan sulfate proteoglycan and calcify to arteriosclerotic nanoplaques (ternary complexes). Nanoplaque formation was measured by ellipsometry, both in vitro on an HS-PG coated hydrophobic silica surface and also in vivo on living human coronary endothelial cells, which had overgrown the silica surface. Reversely, we show with the same techniques that, in dependence on the degree of peroxidation and epitope in concern, oxLDL attacks its molecular receptor and thus can induce degradation of arteriosclerotic plaques and, in a combined action with inflammatory processes, even a plaque rupture. In a previous work, we had found PML-NB, fibrous cap (collagens, proteoglycans) and HSBGF binding sites (e.g., TGF beta 1) up-regulated and NF kappa B down-regulated. With this background knowledge we created a molecular feedback control circuit model where PML-NB functions as regulation centre, fibrous cap as controlled variable, HSBGF binding sites as receptor and NF kappa B as effector. Since NF kappa B is inhibited by one reaction strand in this model and inhibits itself collagen and proteoglycan synthesis in the fibrous cap of the plaque, this double check (disinhibition) causes a stabilization of the fibrous cap through a specially strong collagen and proteoglycan production, which in addition is supported by circulating TGF beta. TGF beta furthers also calcification, so that fibrous cap tensile strength and resistance to shear stress are imparted. This way, a plaque rupture may possibly be averted.
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| 31. |
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| 32. |
- Zhang, Yuning, et al.
(författare)
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Off-Stoichiometric Thiol-Ene Chemistry to Dendritic Nanogel Therapeutics
- 2019
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Ingår i: Advanced Functional Materials. - : Wiley-VCH Verlag. - 1616-301X .- 1616-3028. ; 29:18
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Tidskriftsartikel (refereegranskat)abstract
- A novel platform of dendritic nanogels is herein presented, capitalizing on the self-assembly of allyl-functional polyesters based on dendritic-linear-dendritic amphiphiles followed by simple cross-linking with complementary monomeric thiols via UV initiated off-stoichiometric thiol-ene chemistry. The facile approach enabled multigram creation of allyl reactive nanogel precursors, in the size range of 190–295 nm, being readily available for further modifications to display a number of core functionalities while maintaining the size distribution and characteristics of the master batch. The nanogels are evaluated as carriers of a spread of chemotherapeutics by customizing the core to accommodate each individual cargo. The resulting nanogels are biocompatible, displaying diffusion controlled release of cargo, maintained therapeutic efficacy, and decreased cargo toxic side effects. Finally, the nanogels are found to successfully deliver pharmaceuticals into a 3D pancreatic spheroids tumor model.
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